JP2011125285A - Method for quickly measuring factor causing early flocculation of yeast, and measurement apparatus therefor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method which enables the highly accurate, quick and convenient measurement of a factor causing early flocculation of yeast contained in a brewing material and further enables the quick measurement of a number of specimens. <P>SOLUTION: The method of quickly measuring a factor causing early flocculation of yeast in a brewing material includes: the process (1) of mixing yeast in the late logarithmic growth phase or thereafter with a water-extracted high-molecular fraction prepared from a test material sample in a buffer and suspending the obtained mixture therein; and the process (2) of irradiating the suspension obtained by the process (1) with visible light, photographing a scattered light with a camera device, image-analyzing the obtained image data and thus determining the degree of whiteness of the suspension to thereby determine the degree of sedimentation of the yeast in the suspension. The measurement is performed while controlling the temperature of the suspension obtained by the process (1) at a constant temperature within the range from 20 to 45°C. Besides, when it is desired to measure the factor of high early flocculation activity, the temperature of the suspension is set at that higher than the temperature range. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a method for quickly measuring a yeast early aggregation factor contained in a brewing raw material used for producing fermented malt beverages such as beer, happoshu, and whiskey, and a yeast for brewing raw material using the measuring method The present invention relates to a method for rapid determination of early aggregation. The present invention also relates to a measuring apparatus for continuously and quantitatively measuring the sedimentation of bacterial cells in a suspension.

Related technology

  In brewing fermented malt beverages such as beer, sparkling liquor, and whiskey, the yeast is appropriately aggregated and settled at the end of the main fermentation by the yeast. Yeast can be recovered by this aggregation and sedimentation. However, if the yeast is not properly aggregated and settled at the end of the main fermentation, the recovered amount of yeast will be insufficient. On the other hand, if the yeast is excessively aggregated and settled, fermentation will not proceed. The yeast agglutinates at the end of the fermentation and when the extract is reduced, the yeasts clump together. This aggregation causes the yeast to settle to the bottom of the culture solution. Involvement of the binding of the lectin-like protein on the surface of yeast cells and the mannose sugar chain of yeast mannan has been reported in this agglutination of yeast itself (Appl. Microbiol. Biotechnol 23: 197-205, 2003).

  Thus, in brewing fermented malt beverages and the like, in a normal fermentation process, the aggregation and precipitation of yeast occurs moderately at the end of the main fermentation by yeast and at the time when the extract of the fermentation liquor is reduced. It has been reported that a phenomenon called “early yeast agglomeration phenomenon” (hereinafter sometimes referred to as “early coagulation phenomenon”) is observed in this yeast agglomeration and sedimentation.

  "Early yeast agglomeration phenomenon" is a phenomenon in which yeast agglomerates and settles in the fermentation process of yeast, especially in the latter stage of fermentation, even though the sugars that can be assimilated by yeast remain in the fermentation broth. This early flocculation phenomenon causes the fermentation to stop when yeast agglomerates and settles (Proc. Congr. Eur. Brew. Conv. 26: 53-60, 1997 (non-patent literature). 1)). Therefore, when this phenomenon is observed, fermentation becomes insufficient, the manufactured product becomes out of specification, and a large amount of damage is caused in brewing a fermented malt beverage or the like.

  This early aggregation phenomenon is considered to be caused by high molecular acidic polysaccharides derived from raw wheat and contained in malt (J. Am. Soc. Brew. Chem. 47: 29-34, 1989). And it is known that the factor causing the early aggregation phenomenon may be generated in the wheat making process or may be present in the raw wheat (J. Inst. Brew., 97, 359-366). , 1991; JP-A-10-179190 (Patent Document 1)). Conventionally, in the brewing of fermented malt beverages made from barley, in order to avoid the early agglomeration phenomenon in the fermentation process, the presence of the early agglomeration of malt and barley is confirmed, and the barley malt does not cause the early agglomeration phenomenon It is desirable to select and use these.

  In order to confirm the presence or absence of early cohesiveness such as malt and barley, a method based on a fermentation test has been adopted as a conventional method. Examples of such methods include KY tube fermentation test method developed by T. Nakamura et al. (Proc. Congr. Eur. Brew. Conv. 26: 53-60, 1997 (Non-patent Document 1), M. Jibiki Universal fermentation test (J. Am. Soc. Brew. Chem. 64 (2): 79-85, 2006 (non-patent document 2)), etc. However, all of these methods are fermentation tests. Is a test in which the actual brewing scale (fermentation test scale) is made small, and it is necessary to ferment the wort.In these methods, the wort preparation usually takes 1 day from the progress of fermentation. It takes about 8 days to check for the presence of early aggregation factors in malt and barley, and in the case of wheat before malting, it takes about 7 days to prepare wheat and wort. A long period of time was required to confirm the presence of early aggregation factors.

  In order to shorten the period of the fermentation test for confirming the presence or absence of this early agglomeration, enzyme is added to the test raw wheat and the raw wheat is subjected to enzyme treatment, and the resulting enzyme-treated product or enzyme-treated product is separated. Developed a method to determine the presence or absence of early cohesive factors in raw wheat by adding the obtained polymer fraction to synthetic wort to make a raw material for fermentation test and measuring the turbidity of the raw material for fermentation test after 48 hours (Japanese Patent Laid-Open No. 10-179190 (Patent Document 1)). This method significantly shortened the period for confirming the presence or absence of early aggregation factors. However, this method is also a method based on a fermentation test, and still requires 48 hours of fermentation test.

  International Publication WO2005 / 073394A1 (Patent Document 2) discloses a method called a cuvette method, in which an early coagulation factor extracted from malt and a yeast in the late logarithmic growth are mixed in a cuvette. This is a method of measuring aggregation / sedimentation by absorbance at a specific wavelength and does not require a fermentation test. However, since this method is based on manual analysis, it is not suitable for processing many samples, and there is room for problems in processing accuracy and measurement errors due to differences in the person performing the analysis.

  Japanese Patent Application Laid-Open No. 2009-171955 (Patent Document 3) proposes a method and apparatus that can quickly measure the analysis of multiple samples while utilizing the principle of the cuvette method.

  Since yeast is susceptible to environmental influences, the measurement value may change easily in the analysis using yeast, and errors may occur due to differences in measurement date, measurement time, or analysis. there were. Moreover, the strength of the aggregating ability of yeast is peculiar to the yeast to be used, and it was difficult to control the sensitivity to precoagulant factors. For this reason, environmental factors affecting yeast aggregation can be identified, yeast agglomeration strength can be controlled, and the presence or absence of early aggregation factors in brewing ingredients can be measured more easily and quickly. Development of a method that can be carried out automatically has been desired.

JP 10-179190 A WO2005 / 073394A1 JP 2009-171955 A

Proc. Congr. Eur. Brew. Conv. 26: 53-60, 1997 J. Am. Soc. Brew. Chem. 64 (2): 79 -85, 2006

The present inventors have already obtained the yeast in the late logarithmic growth phase or later and the water-extracted polymer fraction of the test raw material sample such as wheat and malt mixed and suspended in a buffer solution. When measuring the degree of sedimentation of yeast suspended in a suspension, the suspension is irradiated with visible light and the light scattered is photographed with a digital video camera, and the resulting image data is analyzed. In addition, by converting into numerical values as whiteness, the yeast early aggregation factor contained in the raw material for production can be measured in a very short time, and moreover, it has succeeded in simultaneously measuring multiple samples (Japanese Patent Laid-Open No. 2009-171955). (Patent Document 3)). In this study, the present inventors investigated environmental factors that could affect yeast aggregation, and from a wide range of possible factors and parameters, the temperature of the suspension was a specific value. It was discovered that at lower temperatures, the cohesiveness of yeast became stronger and no difference was observed. That is, it has been found that the higher the temperature, the higher the floating property, and the lower the temperature, the lower the floating property. As a result, it has been found that in the measurement method described above, the temperature of the suspension to be measured is set within a specific temperature range, so that the early aggregation factor can be detected accurately and quickly. Furthermore, by setting the temperature of the suspension to a high temperature, the inventors succeeded in preferentially and rapidly detecting an early aggregation factor having high aggregation properties.
The present invention is based on these findings.

  Therefore, an object of the present invention is to provide a method capable of measuring a yeast early aggregation factor contained in a brewing raw material with high accuracy, quickly and simply, and further capable of measuring multiple samples rapidly.

The measurement method according to the present invention is a rapid measurement method of yeast early aggregation factor contained in a brewing raw material,
(1) The yeast in the late logarithmic growth phase or later and the water-extracted polymer fraction prepared from the test raw material sample are mixed and suspended in a buffer solution,
(2) The suspension obtained in step (1) is irradiated with visible light and the scattered light is photographed with a camera device, and the obtained image data is subjected to image analysis, By determining the whiteness, measure the degree of sedimentation of the yeast in the suspension,
Here, the measurement is performed by controlling the temperature of the suspension obtained in the step (1) to a constant temperature within a range of 20 to 45 ° C., and measuring a yeast early aggregation factor having strong precoagulant activity. If desired, the temperature of the suspension is set to a higher temperature within the temperature range.

According to one preferable aspect of the present invention, in the measurement, the temperature of the suspension obtained in the step (1) is controlled to a constant temperature within a range of 30 to 40 ° C.

  According to one preferred embodiment of the present invention, in the method of the present invention, in step (2), the temperature of the suspension is controlled at a constant temperature in the range of 30 to 35 ° C. When it is desired to measure yeast early aggregation factor having strong coagulation activity, the suspension temperature is controlled to a constant temperature higher than 35 ° C.

  According to one more preferred aspect of the present invention, in the method of the present invention, in step (2), the temperature of the suspension is controlled at a constant temperature in the range of 32 to 35 ° C., and measurement is performed. When it is desired to measure a yeast early aggregation factor having a strong precoagulant activity, the suspension temperature is controlled at a constant temperature in the range of 37 to 39 ° C.

  According to one more preferred aspect of the present invention, the method of the present invention comprises a step (2) wherein the suspension temperature is controlled to a constant temperature of 35 ° C. or lower, the suspension temperature is Measurements performed at a constant temperature higher than 35 ° C. are simultaneously performed.

  According to one more preferred aspect of the present invention, in the measurement method of the present invention, controlling to a constant temperature means that the temperature of the suspension is kept within a temperature range of ± 0.5 ° C. during the measurement. To do.

  In the measurement method according to the present invention, preferably, in step (2), the scattered light is photographed with a digital camera device, the obtained image data is subjected to image analysis, and the whiteness of the digitized suspension is determined. Comprising obtaining.

  According to another preferred embodiment of the present invention, the method of the present invention is such that the whiteness of the suspension is 0, the whiteness at the time of quenching is zero, and the yeast immediately after mixing and suspension is uniformly dispersed. This includes numerically setting the whiteness of the turbid liquid as 100.

  According to another preferred embodiment of the present invention, in step (2), the suspension is irradiated with visible light from a light source placed directly under the suspension and scattered in the suspension. The emitted light is photographed from a camera device installed in a horizontal direction with respect to the suspension.

  According to another preferred embodiment of the present invention, in the step (1), a plurality of specimens are mixed and suspended at the same time using a multi-sample shaking apparatus capable of shaking a plurality of specimens at the same time. A suspension is obtained. At this time, more preferably, the degree of sedimentation of yeast in a plurality of suspensions is measured simultaneously.

  According to still another preferred embodiment of the present invention, the yeast used in the step (1) is obtained by culturing the yeast and recovering the yeast in the late logarithmic growth phase or later, or the recovered yeast. Yeast further frozen and stored. More preferably, at this time, the recovered yeast is washed with EDTA.

  According to still another preferred embodiment of the present invention, is the polymer fraction used in the step (1) a polymer fraction prepared by ethanol precipitation of an aqueous extract of a test material sample? Alternatively, it is a polymer fraction obtained by separating a water extract of a test raw material sample by dialysis, ultrafiltration, or gel filtration.

According to another preferred embodiment of the present invention, the polymer fraction used in step (1) is a polymer fraction prepared from a saccharified solution of a test raw material sample.
According to another preferred embodiment of the present invention, when preparing the water-extracted polymer fraction used in step (1), the test raw material sample is treated with an enzyme during extraction.

According to still another preferred embodiment of the present invention, acetate buffer-CaCl ₂ is used as the buffer solution in step (1).

According to still another preferred embodiment of the present invention, a saccharide component selected from the group consisting of glucose, maltose, mannose and a mixture thereof is added to acetate buffer-CaCl ₂ as the buffer solution in step (1). Use the same thing.

According to one preferable aspect of the present invention, the test raw material sample is barley, malt, or barley in the middle of malting.
According to another preferred embodiment of the present invention, the water-extracted polymer fraction of the test raw material sample is a polymer fraction of an extract obtained by extracting the malt pulverized product with water for 30 seconds or more, or It is a polymer fraction of an extract obtained by extracting a barley pulverized product or a barley pulverized product in the middle of wheat making with water for 15 minutes or more.

  The rapid determination method of yeast early aggregation property of the raw material for brewing according to the present invention is characterized by using the rapid measurement method of yeast early aggregation factor according to the present invention. This determination method typically includes a step of determining whether the raw material for brewing is yeast early aggregating ability or not from the result using the rapid measurement method for yeast early aggregation factor according to the present invention. .

  The method for producing malt according to the present invention manages the malt production process by determining the early cohesiveness of the malt raw material, the malt during production, or the produced malt using the method for rapid measurement of yeast early aggregation factor according to the present invention. It is characterized by doing.

  The method for producing a fermented alcoholic beverage according to the present invention is characterized in that the brewing raw material to be used is selected and adjusted by determining the early flocculating property of the brewing raw material using the method for quickly measuring yeast early flocculation factor according to the present invention. And

Furthermore, according to the present invention, there is provided a measuring device for continuously quantitatively measuring the sedimentation of bacterial cells in a suspension,
A plurality of cuvettes or vials for holding a suspension as a specimen are held in a row in a horizontal direction at a predetermined position, and if necessary, the cuvettes or vials are shaken to suspend the cuvettes or vials therein. A sample shaking means,
Temperature control means for controlling the temperature of the suspension in the cuvette or vial to a constant temperature within the range of 20 to 45 ° C .;
A camera device for photographing the cuvettes or vials arranged in a row;
There is provided a measuring device comprising data processing means for analyzing image data taken by the camera device and obtaining numerical data of the whiteness of the suspension.

  According to a preferred aspect of the present invention, in the measuring apparatus of the present invention, the temperature control means controls the temperature of the suspension to a constant temperature within the range of 30 to 40 ° C.

  According to a preferred aspect of the present invention, the sample shaking means includes the temperature control means.

  According to a preferred aspect of the present invention, the temperature control means controls the temperature of the suspension in a part of the plurality of cuvettes or vials to a constant temperature of 35 ° C. or less and in another cuvette or vial. The suspension temperature can be controlled to a constant temperature higher than 35 ° C.

  According to a preferred aspect of the present invention, the sample shaking means has a visible light source directly under the cuvette or vial.

  According to another preferred embodiment of the present invention, the measurement apparatus is configured such that in the specimen shaking means, the cuvette or the vial is shaken for a predetermined time and then allowed to stand for a predetermined time, and photographing by the camera device is performed. In addition, it further includes control means for controlling the sample shaking means and the camera device.

  According to another more preferable aspect of the present invention, in the measurement apparatus, the camera device is a digital video camera.

  According to the preferable aspect of this invention, the said measuring apparatus is used for the rapid measurement of the yeast early aggregation factor contained in a brewing raw material.

  According to a more preferred aspect of the present invention, the measuring device implements the method for quickly measuring a yeast early aggregation factor according to the present invention.

  According to the method for rapid measurement of yeast early aggregation factor of the present invention, when brewing a fermented malt beverage or the like, the presence or absence of an early aggregation factor of the raw material used for brewing is not a fermentation method like the conventional method, but a simple means. Thus, it is possible to measure quickly and accurately, and it is possible to measure multiple samples simultaneously, accurately and efficiently, compared to the conventional cuvette method using absorbance. In the cuvette method, there is no clear rule regarding the method of mixing the yeast and the water-extracted polymer fraction, and there is a possibility of mixing in different ways depending on the person performing the analysis. There was room to produce. In the method according to the present invention, since the processing is simple and multiple samples can be measured simultaneously, it can be said that there is no need to consider such concerns.

  Furthermore, according to the present invention, by setting the temperature of the suspension to be measured within a specific temperature range, the early aggregation factor can be detected more accurately and quickly. In addition, by setting the temperature of the suspension to a high temperature, it is possible to preferentially and more quickly detect an early aggregation factor having high aggregation properties. For this reason, when the presence of an early aggregation factor having high aggregation properties can be expected, the early aggregation factor can be detected more rapidly by adjusting the temperature condition. If such an early aggregation factor having high cohesiveness can be detected more efficiently, the occurrence of damage in the brewing process of a fermented malt beverage or the like can be found earlier. In addition, according to the present invention, when measuring, the temperature of the suspension is set for a plurality of samples in a normal range and a higher range, respectively, and by simultaneously measuring them, early aggregation in the specimen is performed. Factor status can be more qualified and efficiently measured. That is, even when the same early aggregation factor is used, it is possible to easily detect the case where the ratio of the highly aggregating factor is higher or not.

  Furthermore, according to the method of the present invention, it is also possible to automate the processing without much manual measurement of multiple samples. For this reason, the method according to the present invention is extremely useful as a practical method for measuring and judging early cohesiveness of brewing raw materials used in brewing fermented malt beverages and the like. In particular, the ability to measure a large number of specimens quickly, efficiently and accurately is highly expected for application to production on an industrial scale. The same applies to the measuring apparatus according to the present invention.

  In addition, the rapid measurement method of the present invention is not limited to malt or barley before wheat production but also barley immediately after harvesting, barley in the middle of wheat production, or other cereals and extracts used for brewing. As a simple, rapid and reliable method that can be applied to measurement, it can be widely used to determine the presence or absence of early aggregation factors in these brewing raw materials. Furthermore, since the measurement method of the present invention can be measured with a small amount of sample, it can be easily used in brewing fermented malt beverages and the like, and is provided as a practical measurement / judgment method with high accuracy. Is. In addition, since the method for measuring the early aggregation factor of the present invention can accurately grasp the measurement of the early aggregation factor for each raw material, it is possible to determine the early aggregation property for each raw material. . Furthermore, since it is possible to measure the activity of a very small amount of early aggregation factor, it is also possible to measure the early aggregation factor for each fraction divided in each brewing raw material. It can be effectively used for specific purposes.

The figure is a conceptual diagram showing the positional relationship between a camera device, a cuvette, a vial, and a light source. The figure is a conceptual diagram of an apparatus according to the present invention. The figure shows an image 10 minutes after the start of monitoring in Example 1. The figure shows the measurement results in Example 2. The figure is a conceptual diagram of how to divide a vial into five stages for measurement. The figure shows the measurement results in Example 3. The figure shows the measurement results in Example 5.

Detailed description of the invention

Rapid measurement method of yeast early aggregation factor The measurement method according to the present invention, as described above, is a rapid measurement method of yeast early aggregation factor contained in a brewing material,
(1) The yeast in the late logarithmic growth phase or later and the water-extracted polymer fraction prepared from the test raw material sample are mixed and suspended in a buffer solution,
(2) The suspension obtained in step (1) is irradiated with visible light and the scattered light is photographed with a camera device, and the obtained image data is subjected to image analysis, By determining the whiteness, measure the degree of sedimentation of the yeast in the suspension,
Here, the measurement is performed by controlling the temperature of the suspension obtained in the step (1) to a constant temperature within a range of 20 to 45 ° C., and measuring a yeast early aggregation factor having strong precoagulant activity. If desired, the temperature of the suspension is set to a higher temperature within the temperature range.

  In the present invention, the brewing raw material is used for producing fermented malt beverages such as beer, happoshu, and whiskey, and for example, malt or barley before malting, barley immediately after harvesting, or Examples include barley in the middle of wheat making, or other grains and extracts used for brewing. The same thing is mentioned as a test raw material sample, Preferably, the test raw material sample is barley, malt, or barley in the middle of wheat making.

Step (1):
In step (1) in the measurement method of the present invention, the yeast in the late logarithmic growth phase or later and the water-extracted polymer fraction prepared from the test raw material sample are mixed and suspended in a buffer solution. .
In the present invention, as in the cuvette method described in International Publication WO2005 / 073394A1, “yeast of logarithmic growth phase or later” is used for the measurement of yeast early aggregation factor. The reason why “yeast in logarithmic growth phase or later” is used is that yeast begins to acquire an appropriate aggregation ability at this time.

Here, “the yeast in the late logarithmic phase or later” can be prepared, for example, as follows.
Yeast used for brewing is cultured in a commonly used yeast medium to create a yeast growth curve, and yeast that has reached the late stage of logarithmic growth or a later stage (usually started at a culture temperature of 8 ° C) From day 4 to day 5) by centrifugation or the like. The recovered yeast is desirably washed with a chelate compound solution such as 0.1 M EDTA.
The recovered yeast may be used as it is, but the yeast that has been cultured and recovered in advance may be stored frozen by, for example, cryopreserving at −80 ° C. with a 15% glycerol solution. The yeast thus cryopreserved can be prepared by thawing or the like when used in the measurement method of the present invention.

The “water-extracted polymer fraction prepared from the test raw material sample” used in the measurement method of the present invention can be prepared, for example, as follows.
First, a test raw material sample such as barley or malt is pulverized as necessary, followed by water extraction or normal saccharification. When water extraction of a test raw material sample is performed, the test raw material sample is subjected to enzyme treatment by adding α-amylase, β-amylase, β-glucanase, protease or the like according to the method described in JP-A-10-179190. It can be performed. To separate the polymer fraction from the water extract when preparing the water extract polymer fraction of the test raw material sample, ethanol precipitation, for example, adding ethanol to the water extract to a final concentration of 50%, The precipitate can be separated and collected by a method of collecting by centrifugation, a method of fractionating and collecting by dialysis, ultrafiltration, gel filtration, or the like.

In step (1) of the measurement method of the present invention, the yeast and the water-extracted polymer fraction are mixed and suspended in a buffer solution. As the buffer solution used here, it is desirable to use acetate buffer-CaCl ₂ . More preferably, the buffer solution is a solution such as 50 mM acetate buffer (pH 4 to 4.5) -0.1% CaCl ₂ .

Here, for example, the yeast itself may have strong aggregating properties such as those collected in the late stationary phase of growth or yeast collected in a factory or the like. If the yeast to be used is such a highly cohesive yeast, it sinks even when measured with normal malt (non-precipitating malt), and as a result, the non-precipitating property of the raw material May not be measured accurately. For this reason, it suppresses the aggregation based on the yeast when the yeast itself has a strong aggregating property, but has little influence on the aggregation based on the precoagulant factor, so that the measurement noise due to the strongly aggregating yeast is measured. It is desirable to eliminate. Accordingly, the present inventors have further recently, in acetate buffer-CaCl _2, saccharide component (e.g., monosaccharides, disaccharides or mixtures thereof) by using a plus as a buffer solution, to the undesired yeast It succeeded in suppressing appropriately the aggregation noise based on (Example 4). It is considered that this is because the saccharide component has an effect of inhibiting the binding between the yeast and the rapid coagulation factor.

Therefore, according to another preferred embodiment of the present invention, a buffer solution obtained by adding a saccharide component (for example, a monosaccharide, a disaccharide, or a mixture thereof) to acetate buffer-CaCl ₂ is used. Such a saccharide component to be added is preferably selected from the group consisting of glucose, maltose, mannose and mixtures thereof.

  When the saccharide component to be added is maltose, the maltose concentration in the buffer is preferably 0.5 to 1.5% by weight, more preferably 0.8 to 1.2% by weight, particularly preferably about 1% by weight. It has been found that the strength of the inhibitory effect on the binding between yeast and precoagulant factors by saccharide components varies depending on the saccharide. In the case of the saccharides listed above, it has been found that mannose> glucose> maltose in the order of strong inhibitory effect. For this reason, when using mannose and glucose stronger than maltose, it is possible to set the use concentration to a concentration lower than the concentration in the case of maltose described above.

According to a particularly preferred embodiment of the present invention, the buffer solution used is a liquid such as 50mM acetic acid buffer _{(pH4~4.5) -0.1% CaCl 2 -1} % maltose.

  In addition, as described above, in the present invention, it is preferable that the saccharide component is added to the buffer solution as described above when the yeast to be used has strong aggregability. Whether the yeast to be used is highly cohesive can be easily confirmed by whether or not normal malt is settled.

  In addition, mixing and suspension treatment is performed by a conventional method such as stirring or shaking a container such as a cuvette or a vial containing a suspension as a specimen (a mixture of the yeast and the water-extracted polymer fraction). It is done by implementing. In the present invention, preferably, a plurality of specimens are mixed and suspended at the same time using a multi-sample shaking apparatus capable of simultaneously shaking a plurality of specimens to obtain a plurality of suspensions. This is useful for measuring multiple samples simultaneously. Here, as will be described later, the multi-sample shaking device can hold a plurality of cuvettes or vials for containing a suspension as a sample in a row in a horizontal direction at a predetermined position, and if necessary, It is preferable that the cuvette or vial can be shaken to suspend the suspension therein.

Step (2):
In step (2) according to the present invention, light scattered by irradiating the suspension obtained in step (1) with visible light is photographed with a camera device, and the obtained image data is subjected to image analysis. Then, the degree of sedimentation of yeast in the suspension is measured by determining the whiteness of the suspension.

In the conventional cuvette method, the degree of sedimentation of yeast in the suspension was measured by measuring the optical density of the suspension using a spectrophotometer. The light scattered by irradiation with visible light is photographed by a camera device, and the obtained image data is subjected to image analysis. For this reason, it is possible to obtain image data by arranging multiple samples and photographing them with a camera device at the same time, instead of measuring manually one sample at a time as in the conventional method. By analyzing this image data, according to the present invention, multiple samples can be measured simultaneously in a short time.
Furthermore, according to the present invention, since photographing and image analysis can be performed continuously, the measurement method according to the present invention can be performed continuously. For example, it is possible to measure changes over time. is there.

  In the measurement method of the present invention, the suspension is measured by controlling the temperature of the suspension obtained in step (1) to a constant temperature within the range of 20 to 45 ° C. When it is desired to measure a yeast early aggregation factor having strong activity, the temperature of the suspension is set to a higher temperature within the temperature range. By controlling in such a temperature range and performing measurement, it is possible to perform more accurate and quick measurement. In addition, when it is desired to measure a yeast early aggregation factor having a strong early coagulation activity, the desired early aggregation factor can be detected more efficiently by changing the measurement temperature condition higher.

  In the method of the present invention, when the temperature of the suspension to be measured is set within a specific temperature range, the temperature adjusting means is not particularly limited. For example, as described above, the mixing and suspending process in the step (1) is performed using a cuvette or a vial containing a suspension as a specimen (a mixture of the yeast and the water extraction polymer fraction) as described above. The container can be shaken by, for example, shaking. At this time, shaking is performed while keeping the liquid temperature constant, and the temperature inside the cuvette is controlled to control the liquid temperature (suspension of the suspension). Temperature) can be controlled. In addition, for example, the cuvette or vial after shaking can be kept at a desired temperature by a temperature control means (for example, a thermostat) in the measuring device, thereby controlling the temperature of the suspension.

  In the present invention, measurement is performed by controlling the temperature of the suspension obtained in the step (1) to a constant temperature within a range of 20 to 45 ° C. The temperature of the suspension to be controlled is preferably 30 to 40 ° C, more preferably 32 to 39 ° C, and further preferably 32 to 35 ° C. These values are not intended for a specific purpose, such as when you want to preferentially measure highly coagulative early aggregation factors (ie, weak, fast, fast, strong). It can be said that this is a preferable temperature range in the case of detection of all fast-coagulation factors including any precoagulable malt. On the other hand, when trying to detect an early aggregation factor with high aggregability, which is relatively problematic in the brewing process, the temperature is preferably set to 35 to 40 ° C, more preferably 37 to 39 ° C.

  In the above, “measurement is performed while controlling at a constant temperature” means that measurement is performed at a specific temperature within the specified temperature range, even under the described temperature range. Here, the constant temperature means, for example, that the set temperature is kept within a temperature range of, for example, ± 0.8 ° C. (preferably ± 0.5 ° C.) during measurement. By measuring at such a fixed temperature (fixed), more accurate measurement is possible.

  According to one preferred embodiment of the present invention, as described above, in the method of the present invention, in the step (2), the temperature of the suspension is in the range of 30 to 35 ° C. (preferably 32 to 35 ° C.). When measurement is performed while controlling at a constant temperature, and measurement of yeast early aggregation factor having a strong precoagulant activity is desired, the temperature of the suspension is higher than 35 ° C (preferably in the range of 37-39 ° C). ) Measure at a constant temperature. More preferably, in step (2), the temperature of the suspension is controlled at a constant temperature in the range of 32 to 35 ° C., and measurement of yeast early agglutination factor having strong precoagulant activity is desired. In some cases, the suspension temperature is controlled at a constant temperature in the range of 37 to 39 ° C. for measurement. By specifying and measuring temperature conditions in this way, more efficient and quick measurement can be performed.

  According to one more preferable aspect of the present invention, as described above, in the method of the present invention, in the step (2), the temperature of the suspension is 35 ° C. or lower (preferably 30 to 35 ° C., more preferably 32 ° C.). The measurement to be performed at a constant temperature in the range of ˜35 ° C. and the temperature of the suspension to be higher than 35 ° C. (preferably in the range of 35-40 ° C., more preferably in the range of 37-39 ° C.). Perform controlled measurements at the same time. More preferably, in step (2), the measurement is performed by controlling the temperature of the suspension to a constant temperature in the range of 32 to 35 ° C, and the temperature of the suspension is controlled to a constant temperature in the range of 37 to 39 ° C. And the measurement performed at the same time. The state of the early aggregation factor in the specimen can be measured more appropriately and efficiently.

  In the present invention, the light source for irradiating the suspension with visible light can be photographed by a camera device to be described later, and the whiteness can be digitized by image analysis of the photographed image data. Any type of light source may be used. For example, a commercially available white LED light source can be used as the light source. Similarly, the luminous intensity of the light source is not limited as long as the measurement of the present invention is possible, but the specimen can be irradiated with light having a luminous intensity of about 10,000 to 30000 mcd, for example, about 18000 mcd.

  The camera device to be used is not particularly limited as long as the captured image data can be output to the image processing device. For this reason, usually, the camera device is preferably a digital camera, preferably a digital video camera. The number of pixels of the digital camera is not particularly limited as long as the whiteness can be obtained by analyzing the image based on the obtained image data. For example, the number of pixels having 1.3 million pixels or more is preferable.

  Therefore, according to a preferred aspect of the present invention, in step (2), the scattered light is photographed with a digital camera device, the obtained image data is subjected to image analysis, and the whiteness of the digitized suspension is measured. Get.

  Here, the digitized whiteness can be obtained, for example, by analyzing the obtained image data and obtaining the ratio of white to black in the region to be measured as the whiteness. In this case, preferably, the whiteness of the suspension is 0, the whiteness at the time of quenching is 0, and the whiteness of the suspension in which the yeast immediately after mixing / suspension is uniformly dispersed is 100. As a numerical value. In this case, the higher the whiteness, the larger the numerical value is between 0 and 100.

  The image analysis of the image data can be performed by combining a personal computer and commercially available software capable of digitizing the white and black areas in the image.

  According to a preferred embodiment of the present invention, the suspension is irradiated with visible light from a light source placed directly under the suspension. At this time, the light scattered in the suspension is usually photographed from a camera device installed in a horizontal direction with respect to the suspension. By doing so, it is possible to accurately capture the scattered light from the suspended yeast, and this can greatly reduce the problem of processing accuracy and measurement error due to the difference in the person performing the analysis. .

According to one preferred embodiment of the present invention, after mixing and suspending in step (1), the mixture is allowed to stand for a predetermined time if necessary, and then suspended again, and then the degree of sedimentation of yeast in the suspension is measured. . The time for standing here is not particularly limited as long as the yeast in the mixed suspension can react with the precoagulant factor to form an aggregate, and any time may be used. .
The suspension time after standing is not particularly limited as long as the yeast agglomerates in the suspension can be uniformly dispersed, and may be any time. Thus, by providing a fixed standing time and a suspension time, the degree of sedimentation of the yeast becomes clear, and accurate and highly reproducible measurement with little variation becomes possible.

  In the measurement method of the present invention, as described above, the degree of sedimentation of yeast in the suspension is measured by determining the whiteness of the suspension. That is, the larger the degree of sedimentation of yeast, the blacker the whiteness value becomes when the suspension is observed from the side. On the other hand, the lower the sedimentation degree and the less the yeast sedimentation, the whiter the suspension appears and the greater the whiteness value. In this way, the sedimentation degree of yeast can be measured from the value of whiteness.

  According to a more preferred embodiment of the present invention, the sedimentation degree of yeast in a plurality of suspensions is measured simultaneously. At this time, more preferably, the mixing / suspension process in the step (1) is performed simultaneously for multiple samples. As described above, both the step (1) and the step (2) can be performed simultaneously on multiple samples, whereby the measurement method of the present invention can be automated without human intervention.

Measuring device According to the present invention, as described above, a measuring device for continuously quantitatively measuring the sedimentation of bacterial cells in a suspension,
A plurality of cuvettes or vials for holding a suspension as a specimen are held in a row in a horizontal direction at a predetermined position, and if necessary, the cuvettes or vials are shaken to suspend the cuvettes or vials therein. A sample shaking means,
Temperature control means for controlling the temperature of the suspension in the cuvette or vial to a constant temperature within the range of 20 to 45 ° C .;
A camera device for photographing the cuvettes or vials arranged in a row;
There is provided a measuring device comprising data processing means for analyzing image data taken by the camera device and obtaining numerical data of the whiteness of the suspension.

  In FIG. 1, the conceptual diagram which showed the positional relationship of a camera apparatus, a cuvette or a vial, and a light source was shown. FIG. 2 shows a conceptual diagram of an apparatus according to the present invention.

  Here, examples of the microbial cells include yeast and mold, and yeast is preferred. The measurement method of the present invention can also be applied to all organisms that can aggregate and settle.

  The specimen shaking means holds a plurality of cuvettes or vials for containing a suspension as a specimen in a row in a horizontal direction at a predetermined position, and shakes the cuvettes or vials as necessary. Thus, the suspension therein can be suspended.

  Here, a plurality of (for example, 11 to 12) cuvettes or vials are held in a row in a horizontal direction at a predetermined position means that a plurality of cuvettes or vials can be photographed simultaneously by a camera device. The cuvette or vial is held at a height or position (position relative to the camera device), and is usually arranged in a line perpendicular to the direction in which the camera device is photographed so that it can be photographed simultaneously. . Therefore, the position of the cuvette or vial can be set as appropriate within the range of this purpose. In this case, the height and position of cuvettes or vials, how they are arranged, and the positional relationship with the camera device may be appropriately changed as long as multiple samples can be simultaneously photographed by the camera device.

  In addition, in the specimen shaking means, if necessary, the cuvette or vial can be shaken to suspend the suspension therein. The suspension is obtained by shaking the cuvette or vial held in place. The apparatus is not particularly limited as long as it is a device that can suspend liquid, for example, a multi-sample shaking device that can fix a plurality of cuvettes or vials in parallel (in a row) and shake them while inverting them. Preferably, such a multi-sample shaking apparatus has variable shaking speed, shaking time, and waiting time, and can be set as appropriate. Such control can be performed using a separate control device. By precisely controlling the shaking speed, shaking time, and waiting time with a control device or the like, it is possible to eliminate measurement errors due to differences in the analysts. As a control apparatus, you may use a commercial item, for example.

  The measuring apparatus of the present invention comprises temperature control means for controlling the temperature of the suspension in the cuvette or vial to a constant temperature within the range of 20 to 45 ° C. (preferably 30 to 40 ° C.). More preferably, the temperature control means controls the temperature of the suspension in a part of the plurality of cuvettes or vials to a constant temperature of 35 ° C. or less, and the suspension in another cuvette or vial. The temperature can be controlled to a constant temperature higher than 35 ° C.

  More specifically, in the measuring apparatus of the present invention, the temperature control means can adjust the internal temperature of the cuvette or the like to 20 to 45 ° C. in order to keep the temperature of the suspension constant. Within a temperature range of 5 ° C.). In the present invention, the temperature inside the cuvette is such that the temperature inside the cuvette is 20 to 45 ° C. (for example, preferably 30 to 40 ° C., more preferably 30 to 35 ° C., more preferably 32 to 35 ° C.). Can be controlled. Increasing the temperature inside the cuvette weakens the cohesiveness (decreases sensitivity to precoagulant factors), and decreases the temperature increases the cohesiveness (increased sensitivity to precoagulant factors). For this reason, it is good to change temperature up and down according to the objective using these.

  As a specific example of the temperature control means, the temperature is controlled by blowing air whose temperature is adjusted by the heating device and the cooling device into the device, or the temperature inside the device is directly controlled by the heating device and the cooling device. And what to do.

  According to a preferred aspect of the present invention, the sample shaking means includes this temperature control means.

More preferably, in the sample shaking means, the sample shaking means and the camera device are arranged such that the sample shaking means causes the cuvette or the vial to shake for a predetermined time, and then is allowed to stand for a predetermined time and photographing is performed by the camera device. And a control means for controlling the above. In this way, by controlling operations from mixing, suspension, standing, and camera photography, measurement can be automated, and errors that can be caused by different analysts can be almost eliminated. Furthermore, it becomes possible to find and set a more optimal measurement condition.
As an example of such control, for example, in the case of a glass vial, the condition of mixing 2 minutes → stand (standing) 15 minutes or longer → mixing 2 minutes → measurement (photographing) can be mentioned. The condition of mixing for 7 minutes, standing (standing) for 2 minutes, mixing for 7 minutes, and measurement (photographing) can be mentioned.

  According to a preferred aspect of the present invention, the specimen shaking means has a visible light source directly under the cuvette or vial.

  In the present invention, the data processing means is capable of obtaining numerical data of the whiteness of the suspension by analyzing the image data captured by the camera device. An example of such processing means is a personal computer. In this case, the personal computer may be used in combination with software that can perform image analysis of the obtained image data, for example, commercially available software that can quantify the white and black areas in the image. preferable.

  In this specification, the expression of a value using “about” and “degree” includes a variation in a value that can be allowed by those skilled in the art to achieve the purpose by setting the value. Meaning. For example, it means that a variation within 20%, preferably within 10%, more preferably within 5% of a predetermined value or range can be tolerated.

  The present invention will be described in detail by the following examples, but the present invention is not limited thereto.

experimental method
1) Yeast culture The pre-cultured solution cultured in a stationary culture at 20 ° C. for 3 days in a YEPG medium (1% Yeast Extract, 2% Bactopepton, 7.5% Maltose, 2.5% Glucose) The solution was added to a cooled YEPG medium (usually 100 ml in a 250 ml medium bottle) at a concentration of 1.5%, and cultured at 8 ° C. while stirring with a stirrer. During culture, OD600 is measured over time, and a yeast growth curve is prepared. In the logarithmic growth phase, the logarithmic growth late stage (4 days after the start of culture), the early stationary phase (5 days 10 days after the start of culture) Eyes) can be used.

2) Preparation of test yeast The yeast in the early stationary phase after completion of the culture was recovered by centrifugation at 3000 rpm for 5 minutes (4 ° C). The recovered yeast is suspended and washed with cold water and similarly collected by centrifugation, suspended twice with cold 100 mM EDTA (pH 8.0) and twice with cold water, and suspended in 20 ml of cold water. It was. The yeast concentration was measured by diluting the yeast solution 1/200 and measuring OD600. Moreover, after performing washing | cleaning by cold water twice, after suspending in a 15% glycerol solution and measuring yeast concentration similarly by measuring OD600 of a yeast solution, it was made to freeze at -80 degreeC.

3) Preparation of malt water extraction polymer fraction 120 ml of water was added to 20 g of each of the following fast-coagulated malt and non-precipitated malt (normal malt), and the mixture was stirred with a stirrer for 10 minutes. After stirring, unnecessary fractions were removed by centrifugation at 8718 g for 10 minutes at 4 ° C., and the supernatant was collected. The supernatant was further filtered to remove the insoluble fraction, and an equal amount of ethanol was mixed. After allowing to stand at room temperature for 10 minutes, the precipitate fraction was collected by centrifugation at 11387 g for 10 minutes at 4 ° C. The precipitate fraction was suspended in 10 ml of hot water to obtain a measurement sample.
Fast-setting malt 1-3: Sample for malt evaluation manufactured by Kirin Brewery Co., Ltd. Normal malt 1-3: Sample for malt evaluation manufactured by Kirin Brewery Co., Ltd.

Example 1: Identification of fast-coagulating malt A water solution was prepared by adding water to the freeze-dried yeast obtained in 2) above, and having a predetermined concentration (OD600 of about 200), while obtained in 3) above. The obtained malt water extraction polymer fractions (fast-coagulating malts 1 to 3 and normal malts 1 to 3) were prepared.
Next, 0.075 ml of the yeast solution and 0.385 ml of the malt water extraction polymer fraction are suspended in 1.5 ml of 50 mM sodium acetate (pH 4.8) and 6 μl of 50% calcium chloride so that the total amount becomes 3 ml. Prepared with water. Each of the obtained solutions was put into a plastic cuvette (10 × 10 × 45 mm), and the upper part was sealed with parafilm.
These were prepared and placed in a folder in a multi-sample shaking apparatus. After shaking for 7 minutes → standing for 2 minutes → penetrating for 7 minutes, sedimentation was monitored under the following conditions.

The multi-sample shaking device to be used is one that can fix eleven cuvettes in parallel as a shaking device part and can be shaken while inverting.
As a light source, a white LED light source (manufactured by Nichia Corporation, NSPW500BS) (AC100V / 3A) was used.
As a control device for the shaking device, KV-16AT manufactured by Keyence Corporation was used.
As a camera device, a digital video camera (ArtCam Co., Ltd., ARTCAM-130MI (1.5 million pixels)) was used.
A commercial computer (OS: Windows XP) was used as the personal computer, and a commercially available precipitation rate analysis software was used.
All measurements were performed at room temperature.

The result was as shown in FIG. The figure is an image 10 minutes after the start of monitoring.
In the figure, normal malt 1, fast-coagulated malt 1, fast-coagulated malt 2, normal malt 2, early-coagulated malt 3, and normal malt 3 are shown in duplicate from the right.
Table 1 below shows numerical values of whiteness obtained by image analysis of the image data shown in the figure.

Example 2: Examination of measurement time A sample of precoagulable malt 1 was prepared in the same manner as in Example 1 above. These were considered according to Example 1 by dividing the vial into five stages (FIG. 5), and monitoring was performed at each stage, and the whiteness was measured over time from time 0 to 30 minutes.

The result was as shown in FIG. In the drawing, series 2 to 6 represent measurement results through windows divided into five stages, and series 2 to 6 mean results at the window positions from the bottom in order.
As a result of examining the measurement time, it was found that in all stages, the turbidity decreased significantly by the first 3 minutes, but was relatively stable until 30 minutes thereafter.

Example 3: Examination of measurement position First, samples of early setting malt and non-early setting malt were prepared in the same manner as in Example 1. Next, in order to examine which position of the vial or cuvette has the highest accuracy, the vial was divided into five stages (FIG. 5), and the turbidity was compared for each area of each stage.

  The result was as shown in FIG. The figure shows the turbidity after standing for 4 minutes. In the figure, the results of dividing each vial into five areas are shown in order. In any vial, it was possible to determine early setting malt and non-precipitating malt by measuring in five areas.

Example 4: Examination of buffer solution The yeast was cultured in the same manner as in the section of “1) Yeast culture” in the “Experimental method” described above. The following was used as follows:
Yeast 1: cultured for 7 days from the start of culture,
Yeast 2: cultured for 16 hours 6 days from the start of culture.
In addition, as a control, cultured yeast in the early stationary phase (cultured for 10 hours for 5 days from the start of culture) was used as “normal yeast” in the following.
Water was added to each of the obtained cultured yeast samples to prepare a yeast solution having a predetermined concentration (OD600 is about 200).

  “Normal malt 1” and “fast-setting malt 1” were prepared as malt samples in the same manner as in “3) Preparation of malt water-extracted polymer fraction” in “Experimental method” described above.

  Other than using the prepared yeast solution, using “normal malt 1” and “fast-coagulating malt 1” as the malt water extraction polymer fraction, and further using the following conditions as a buffer: The test was conducted in the same manner as in Example 1 to determine the whiteness for each case.

  The results were as shown in Table 2.

As is apparent from the results in the case where no saccharide component is added, the use of yeast with strong aggregability (ie, yeast 1 and yeast 2) is due to the fact that normal malt is used due to the agglutination of the yeast itself. Also, the whiteness decreased.
When 1% maltose was added to the buffer solution when yeast having strong cohesiveness was used, a clear difference was observed in the whiteness value between normal malt and fast-coagulated malt. As a result of such a clear difference, it has become clear that it is possible to determine whether the malt used is fast-coagulating. In the above table, such a difference was remarkable when the sugar component used was maltose and the concentration was 1% by weight.

Example 5: Examination of measurement temperature In the same manner as in the section of “3) Preparation of malt water extraction polymer fraction” in “Experimental method” described above, “normal malt 1” (normal malt) and “ Precocious malt 1 ”(precocious malt) was prepared.
Moreover, based on the wort residual sugar degree when wort is actually fermented in the fermentation test, the higher one compared with the case of normal malt is collected as “Kohaya coagulation malt” (Kogaya coagulation malt 1 and 2), Moreover, what was low compared with the case of normal malt was extract | collected as "weak early setting malt" (all are samples for malt evaluation by Kirin Brewery Co., Ltd.). These were made into malt samples in the same manner as in the section of “3) Preparation of malt water extraction polymer fraction” in the “Experimental method” described above, and “Kogaya coagulation malt 1” (Kogaya coagulation malt), “ It was prepared as “Kohaya early malt 2” (Kohaya early malt) and “Kayahaya malt” (weakly early malt).

  In addition to “normal malt” and “fast-coagulating malt” as a high molecular fraction extracted from the malt water using the prepared yeast liquid, “Kohaya-coagulated malt 1”, “Kokaya-coagulated malt 2”, “weak” The test was carried out in the same manner as in “Example 1” except that “fast-cooking malt” was used and the conditions described in Table 3 were used as measurement temperature conditions, and the whiteness for each sample was determined. Asked.

  Here, the measuring device used was such that the internal temperature could be adjusted to 22 to 40 ° C. in order to keep the suspension temperature constant, and control was possible at ± 0.5 ° C. . In the experiment, the temperature inside the cuvette was controlled so that the temperature inside the cuvette was constant at each temperature shown in the following table of 32 to 35 ° C.

  The results were as shown in Table 3 and FIG.

Claims (31)

A method for quickly measuring a yeast early aggregation factor contained in a brewing material,
(1) The yeast in the late logarithmic growth phase or later and the water-extracted polymer fraction prepared from the test raw material sample are mixed and suspended in a buffer solution,
(2) The suspension obtained in step (1) is irradiated with visible light and the scattered light is photographed with a camera device, and the obtained image data is subjected to image analysis, By determining the whiteness, measure the degree of sedimentation of the yeast in the suspension,
Here, the measurement is performed by controlling the temperature of the suspension obtained in the step (1) to a constant temperature within a range of 20 to 45 ° C., and measuring a yeast early aggregation factor having strong precoagulant activity. If desired, the measuring method is characterized in that the temperature of the suspension is set to a higher temperature within the temperature range.   The method according to claim 1, wherein in the measurement, the temperature of the suspension obtained in the step (1) is controlled to a constant temperature within a range of 30 to 40 ° C.   In the step (2), when the temperature of the suspension is controlled at a constant temperature in the range of 30 to 35 ° C. and measurement of yeast early agglutination factor having a strong precoagulant activity is desired, The method according to claim 1 or 2, wherein the measurement is performed while controlling the temperature of the suspension at a constant temperature higher than 35 ° C.   In the step (2), when the temperature of the suspension is controlled at a constant temperature in the range of 32 to 35 ° C. and measurement of yeast early agglutination factor having a strong precoagulant activity is desired, The method according to any one of claims 1 to 3, wherein the measurement is performed while controlling the temperature of the suspension at a constant temperature in a range of 37 to 39 ° C.   In the step (2), the measurement performed by controlling the temperature of the suspension to a constant temperature of 35 ° C. or lower and the measurement performed by controlling the temperature of the suspension to a constant temperature higher than 35 ° C. are performed simultaneously. The method as described in any one of Claims 1-4.   Control according to any one of claims 1 to 5, wherein controlling to a constant temperature means keeping the temperature of the suspension within a temperature range of ± 0.5 ° C during the measurement.   In the step (2), the scattered light is photographed with a digital camera device, and the obtained image data is subjected to image analysis to obtain a quantified whiteness of the suspension. The method according to one item.   The whiteness of the suspension is quantified by setting the whiteness at the time of quenching to 0 and the whiteness of the suspension in which the yeast immediately after mixing and suspension is uniformly dispersed as 100. The method according to any one of 7 above.   In step (2), the suspension is irradiated with visible light from a light source placed directly under the suspension, and the light scattered in the suspension is horizontally directed to the suspension. The method according to any one of claims 1 to 8, wherein an image is taken from an installed camera device.   In the step (1), a plurality of specimens are mixed and suspended at the same time using a multi-sample shaking apparatus capable of simultaneously shaking a plurality of specimens to obtain a plurality of suspensions. The method according to claim 1.   The method according to claim 10, wherein the degree of sedimentation of yeast in a plurality of suspensions is measured simultaneously.   The yeast used in the step (1) is a yeast obtained by culturing yeast and recovering yeast in the late logarithmic growth phase or later, or the recovered yeast is further cryopreserved. 12. The method according to any one of 11 above.   The polymer fraction used in step (1) is a polymer fraction prepared by ethanol precipitation of a water extract of a test raw material sample, or a water extract of a test raw material sample is dialyzed, The method according to any one of claims 1 to 12, which is a polymer fraction separated by ultrafiltration or gel filtration.   The method according to any one of claims 1 to 13, wherein the polymer fraction used in step (1) is a polymer fraction prepared from a saccharified solution of a test raw material sample.   The method according to any one of claims 1 to 14, wherein, in preparing the water-extracted polymer fraction used in step (1), the test raw material sample is subjected to an enzyme treatment during extraction. As the buffer solution of step (1), using the acetate buffer-CaCl _2, The method according to any one of claims 1 to 15. The buffer solution used in step (1) is one obtained by adding a saccharide component selected from the group consisting of glucose, maltose, mannose and a mixture thereof to acetate buffer-CaCl ₂ . The method according to one item.   The method according to any one of claims 1 to 17, wherein the test raw material sample is barley, malt, or barley in the middle of malting.   The water-extracted polymer fraction of the test raw material sample is the polymer fraction of the extract obtained by extracting the malt pulverized product with water for 30 seconds or more, or the barley pulverized product or the barley pulverized product in the middle of malting The method according to any one of claims 1 to 18, which is a polymer fraction of an extract extracted with water for 15 minutes or more.   A method for quickly judging yeast early aggregability of a raw material for brewing, wherein the method according to any one of claims 1 to 19 is used.   The malt production process is managed by determining the early cohesiveness of malt raw material, malt during production, or production malt using the method according to any one of claims 1 to 19. , A method for producing malt.   Production of a fermented alcoholic beverage characterized by selecting and adjusting the brewing raw material to be used by determining the early cohesiveness of the brewing raw material by using the method according to any one of claims 1 to 19. Method. A measurement device for continuously and quantitatively measuring sedimentation of bacterial cells in a suspension,
A plurality of cuvettes or vials for holding a suspension as a specimen are held in a row in a horizontal direction at a predetermined position, and if necessary, the cuvettes or vials are shaken to suspend the cuvettes or vials therein. A sample shaking means,
Temperature control means for controlling the temperature of the suspension in the cuvette or vial to a constant temperature within the range of 20 to 45 ° C .;
A camera device for photographing the cuvettes or vials arranged in a row;
A measuring apparatus comprising: data processing means for analyzing image data captured by the camera device to obtain numerical data of the whiteness of the suspension.   The measuring device according to claim 23, wherein the temperature control means controls the temperature of the suspension to a constant temperature within a range of 30 to 40 ° C.   The measurement apparatus according to claim 23 or 24, wherein the sample shaking means includes the temperature control means.   The temperature control means controls the temperature of the suspension in a part of the plurality of cuvettes or vials to a constant temperature of 35 ° C. or lower, and the temperature of the suspension in the other cuvettes or vials from 35 ° C. The measuring device according to any one of claims 23 to 25, which can be controlled to a high constant temperature.   27. The measuring apparatus according to any one of claims 23 to 26, wherein the specimen shaking means includes a visible light source directly below the cuvette or vial.   The sample shaking means further includes a control means for controlling the sample shaking means and the camera device such that the cuvette or the vial is shaken for a predetermined time and then allowed to stand for a predetermined time, and photographing by the camera device is performed. The measurement apparatus according to any one of claims 23 to 27.   The measurement apparatus according to any one of claims 23 to 28, wherein the camera apparatus is a digital video camera.   The measuring device according to any one of claims 23 to 29, which is used for rapid measurement of a yeast early aggregation factor contained in a brewing raw material.   The measurement apparatus according to any one of claims 23 to 30, wherein the method according to any one of claims 1 to 19 is performed.